M.J. Welsh and A.E. Smith, RAC Application 
Ml 3-1. These studies will be performed on the cytospin preparations from nasal brushing 
and on the biopsy specimen. 
g. Transepithelial electrical potential difference across the nasal epithelium. 
Purpose : To assess the electrophysiological consequences of treatment. Patients with CF 
have an abnormally increased transepithelial electrical potential difference across the nasal 
epithelium (103,104). In addition, the response of the electrical potential difference to 
several agents that regulate transport is abnormal. The abnormalities in patients with CF 
are well characterized and relatively easy to measure. Correction of the abnormality would 
indicate that administration of the recombinant virus has corrected the CF electrolyte 
transport defect in a localized area of nasal epithelium. 
Methods : Measurement of the nasal PD is safe, easy, and not invasive (103,104). The 
reference bridge consists of a polyethylene PE 240 tubing filled with 3 M KC1 with 4% 
agar placed under an EKG patch (3-M) on the forearm. The reference bridge is connected 
through a calomel cell to a voltmeter (602 electrometer, Keithly Instruments). The 
exploring bridge is a small PE 50 tubing perfused with saline solution (0.2 ml/min, Harvard 
Compact Infusion Pump) which is connected to a 3 M KC1 bridge and then a calomel cell. 
The electrical potential difference between the exploring and reference electrodes is 
measured by a voltmeter. Output of the voltmeter is connected to a strip chart recording 
(Servocorder SR 6253 Datamark). The exploring electrode (the PE50 tubing) will be used 
to gently touch the site of virus application under direct vision. After a five minute 
baseline, the infusion solution will be changed to one consisting of Ringer's solution 
containing either 100 pM terbutaline, 100 pM amiloride, 100 pM adenosine, and/or saline 
in which die chloride concentration has been reduced by replacement with gluconate. The 
entire procedure will take between 15-30 minutes. Treated and non-treated adjacent 
mucosa will be analyzed for each condition. 
h. Mucosal fluid for adenovirus antibody. 
Purpose: To test for the presence on the mucosal surface of IgA or IgG antibody to the 
recombinant adenoviruses and to test for neutralizing antibodies. 
Methods: Before administration of the virus and at one month after we will obtain a 
washing of the nasal surface to test for the presence of specific and neutralizing antibody. 
5. Evaluation after discharge. 
After discharge from the Clinical Research Center, patients will be evaluated at weekly or 
every other week intervals for 6 weeks. We will continue to make the same assessments as 
described above. The data may be useful in assessing the persistence of expression of 
CFTR. 
B.3.a. Will cells (eg. bone marrow cells) be removed from patients and treated ex vivo? If 
so. what kinds of cells will be removed from the patients, how many, how often, and at 
what intervals? 
No. 
B.3.b. Will patients be treated to eliminate or reduce the number of cells containing 
malfunctioning genes (eg. through radiation or chemotherapy)? 
No. 
Recombinant DNA Research, Volume 16 
[913] 
